5. Test no. 442A: skin sensitization. Local lymph node stimulation test (DA)

The Local Lymph Node Stimulation Assay: DA (ELGL: DA) is a non-radioactive variant of the ELGL method, designed to identify sensitizing chemicals and measure the lymphocyte proliferation they induce in auricular lymph nodes. This method, described in mice (CBA/J strain), is based on bioluminescence quantification of adenosine triphosphate (ATP) content, an indicator of this proliferation.

At least four animals are used per dose group, with a minimum of three concentrations of the test substance, plus a negative control group treated only with the vehicle, and if necessary a positive control. The experimental protocol lasts eight days. ATP content must be quantified within 30 minutes of euthanasia. The procedure applied, from lymph node excision to ATP determination, must be identical for each animal and must not exceed 20 minutes.

The luciferin/luciferase...